Functional analysis of HCV core protein using HCV core transgenic mouse

Research Project

Project/Area Number	11670476
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Single-year Grants
Section	一般
Research Field	Gastroenterology
Research Institution	Chiba University
Principal Investigator	IMAZEKI Fumio Chiba University, School of Medicine, Lecturer, 医学部, 講師 (40223325)
Co-Investigator(Kenkyū-buntansha)	TOKUHISA Takeshi Chiba University, Graduate School of Medicine, Professor, 大学院・医学研究科, 教授 (20134364) HONDA Arata Chiba University, University Hospital, Researcher, 医学部, 医員 YOKOSUKA Osamu Chiba University, School of Medicine, Lecturer, 医学部, 講師 (90182691)
Project Period (FY)	1999 – 2000
Project Status	Completed (Fiscal Year 2000)
Budget Amount *help	¥3,600,000 (Direct Cost: ¥3,600,000) Fiscal Year 2000: ¥1,700,000 (Direct Cost: ¥1,700,000) Fiscal Year 1999: ¥1,900,000 (Direct Cost: ¥1,900,000)
Keywords	HCV core protein / transgenic mouse / suppression subtractive hybridization / cDNA array / DNA chip / mRNA / トランスジェニックマウス / C型肝炎ウイルスコア蛋白 / サブトラクション法 / 肝細胞癌 / DNAチップ / C型慢性肝炎 / Northern blotting / C型肝炎ウイルス / HCVコアタンパク / Fasシグナル伝達系 / 細胞周期 / 細胞増殖
Research Abstract	We analyzed comprihensively the difference of mRNAs expressed in the livers of HCV core transgenic mouse and wild mouse in order to clarify the function of HCV core protein in vivo by the method of subtraction and cDNA microarray. We have got fifty clones of HCV core transgenic mouse subtracted from wild one and 44 clones of wild mouse subtracted from HCV core transgenic one by the method of suppression subtractive hybridization. Sequence analysis of these clones showed not any genes associated with inflammation or carcinogenesis, and the intensity of expression of these genes between transgenic and wild ones was within two times. Next, we analyzed the difference of expression of about 1000 genes between transgenic and wild mice by cDNA array and DNA chip, respectively. Both methods revealed several genes expressed more than two times in transgenic mouse, and these genes seem also not associated with inflammation or carcinogenesis. We will analyze the significance of these genes from now. Expression of HCV core mRNA was ascertained in the transgenic mouse used in this study by northern blotting and the amount of mRNA was estimated to be around 2 pg/μgRNA in the liver. HCV core protein was not analyzed in the taransgenic mouse used in this study, but previous results showed the expression of HCV core protein by western blotting. Therefore HCV core protein might have little effect on the expression of genes in this transgenic mouse in vivo.